Abstract
The widespread use of β-lactam antibiotics has given rise to a dramatic increase in clinically-relevant β-lactamases. Understanding the structure/function relation in these variants is essential to better address the ever-growing incidence of antibiotic resistance. We previously reported the backbone resonance assignments of a chimeric protein constituted of segments of the class A β-lactamases TEM-1 and PSE-4 (Morin et al. in Biomol NMR Assign 4:127–130, 2010. doi:10.1007/s12104-010-9227-8). That chimera, cTEM17m, held 17 amino acid substitutions relative to TEM-1 β-lactamase, resulting in a well-folded and fully functional protein with increased dynamics. Here we report the 1H, 13C and 15N backbone resonance assignments of chimera cTEM-19m, which includes 19 substitutions and exhibits increased active-site perturbation, as well as one of its deconvoluted variants, as the first step in the analysis of their dynamic behaviours.
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Acknowledgments
The authors thank Michelle M. Meyer and Frances H. Arnold for providing the original cTEM-19m construct; Tara Sprules and Sameer Al-Abdul-Wahid from the Québec/Eastern Canada High Field NMR Facility for NMR technical assistance; and the Concordia University BIOFINS platform for access to the circular dichroism apparatus. This work was supported by Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grants RGPIN 227853 and 402623 (to J.N.P and N.D, respectively), in addition to a grant from the National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health (NIH) under Award No. R01GM105978 (to N.D.). S.G. is the recipient of a FRQNT Graduate Scholarship and D.G. is the recipient of an NSERC Alexander Graham Bell Canada Graduate Scholarship. N.D. holds a Fonds de Recherche Québec – Santé (FRQS) Research Scholar Junior 1 Career Award.
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Gobeil, S.M.C., Gagné, D., Doucet, N. et al. 15N, 13C and 1H backbone resonance assignments of an artificially engineered TEM-1/PSE-4 class A β-lactamase chimera and its deconvoluted mutant. Biomol NMR Assign 10, 93–99 (2016). https://doi.org/10.1007/s12104-015-9645-8
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DOI: https://doi.org/10.1007/s12104-015-9645-8